Human IL-2 Antibody

R&D Systems | Catalog # AF-202-NA

R&D Systems
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, ELISA, Neutralization, Immunocytochemistry

Cited:

Neutralization, ELISA Capture, ELISA Detection

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IL-2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human IL-2
Ala21-Thr153
Accession # P60568

Specificity

Detects human IL-2 in direct ELISAs and Western blots. In direct ELISAs, greater than 50% cross-reactivity with recombinant rabbit IL-2 is observed and less than 20% cross-reactivity with recombinant bovine IL-2, recombinant mouse IL-2 and recombinant rat IL-2 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human IL-2 Antibody

Detection of Human IL-2 antibody by Western Blot.

Detection of Human IL‑2 by Western Blot.

Western blot shows lysates of monensin treated human peripheral blood mononuclear cells (PBMCs) with no additional treatment (-) or additionally treated (+) with 0.5 μg/mL calcium ionomycin (Iono) and 50 ng/mL PMA overnight. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for IL-2 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
IL-2 antibody in Human PBMCs by Immunocytochemistry (ICC).

IL‑2 in Human PBMCs.

IL-2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PMA, ionomyocin, and monensin using Goat Anti-Human IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; (NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cell Proliferation Induced by IL‑2 and Neutralization by Human IL‑2 Antibody.

Cell Proliferation Induced by IL‑2 and Neutralization by Human IL‑2 Antibody.

Recombinant Human IL-2 (202-IL) stimulates proliferation in the CTLL-2 mouse cytotoxic T cell line in a dose-dependent manner (orange line) as measured by Resazurin (AR002). Proliferation elicited by Recombinant Human IL-2 (2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA). The ND50 is typically ≤ 0.15 µg/mL.
Human IL-2 ELISA Standard Curve

Human IL-2 ELISA Standard Curve

Recombinant Human IL‑2 (Catalog # 202-IL) was serially diluted and captured by Mouse Anti-Human/Primate IL‑2 Monoclonal Antibody (Catalog # MAB602) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Human IL-2 Antibody

Application
Recommended Usage

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human IL‑2 Monoclonal Antibody (Catalog # MAB2021).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human IL-2 DuoSet ELISA Kit (Catalog # DY202) for convenient development of a sandwich ELISA or the Human IL-2 Quantikine ELISA Kit (Catalog # D2050) for a complete optimized ELISA.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells stimulated with PMA, ionomycin, and monensin

Western Blot

0.5 µg/mL
Sample:

Human peripheral blood mononuclear cells (PBMCs) treated with monensin, 0.5ug/mL calcium ionomycin and 50ng/mL PMA overnight

Neutralization

Measured by its ability to neutralize IL‑2-induced proliferation in the CTLL‑2 mouse cytotoxic T cell line. Gearing, A.J.H. and C.B. Bird (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 276. The Neutralization Dose (ND50) is typically ≤0.15 µg/mL in the presence of 2 ng/mL Recombinant Human IL‑2.

Reviewed Applications

Read 2 reviews rated 5 using AF-202-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IL-2

Interleukin-2 (IL-2) is a O-glycosylated, four alpha -helix bundle cytokine that has potent stimulatory activity for antigen-activated T cells. It is expressed by CD4+ and CD8+ T cells, gamma delta T cells, B cells, dendritic cells, and eosinophils (1 - 3). Mature human IL-2 shares 56% and 66% aa sequence identity with mouse and rat IL-2, respectively. Human and mouse IL-2 exhibit cross-species activity (4). The receptor for IL-2 consists of three subunits that are present on the cell surface in varying preformed complexes (5 - 7). The 55 kDa IL-2 R alpha is specific for IL-2 and binds with low affinity. The 75 kDa IL-2 R beta, which is also a component of the IL-15 receptor, binds IL-2 with intermediate affinity. The 64 kDa common gamma chain gamma c/IL-2 R gamma, which is shared with the receptors for IL-4, -7, -9, -15, and -21, does not independently interact with IL-2. Upon ligand binding, signal transduction is performed by both IL-2 R beta and gamma c. IL-2 is best known for its autocrine and paracrine activity on T cells. It drives resting T cells to proliferate and induces IL-2 and IL-2 R alpha synthesis (1, 2). It contributes to T cell homeostasis by promoting the Fas-induced death of naïve CD4+ T cells but not activated CD4+ memory lymphocytes (8). IL-2 plays a central role in the expansion and maintenance of regulatory T cells, although it inhibits the development of Th17 polarized cells (9 - 11). Thus, IL-2 may be a key cytokine in the natural suppression of autoimmunity (12, 13).

References

  1. Ma, A. et al. (2006) Annu. Rev. Immunol. 24:657.
  2. Gaffen, S.L. and K.D. Liu (2004) Cytokine 28:109.
  3. Taniguchi, T. et al. (1983) Nature 302:305.
  4. Mosmann, T.R. et al. (1987) J. Immunol. 138:1813.
  5. Liparoto, S.F. et al. (2002) Biochemistry 41:2543.
  6. Wang, X. et al. (2005) Science 310:1159.
  7. Bodnar, A. et al. (2008) Immunol. Lett. 116:117.
  8. Jaleco, S. et al. (2003) J. Immunol. 171:61.
  9. Malek, T.R. (2003) J. Leukoc. Biol. 74:961.
  10. Laurence, A. et al. (2007) Immunity 26:371.
  11. Kryczek, I. et al. (2007) J. Immunol. 178:6730.
  12. Afzali, B. et al. (2007) Clin. Exp. Immunol. 148:32.
  13. Fehervari, Z. et al. (2006) Trends Immunol. 27:109.

Long Name

Interleukin 2

Alternate Names

Aldesleukin, IL2, Proleukin, TCGF

Entrez Gene IDs

3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)

Gene Symbol

IL2

UniProt

P60568

Additional IL-2 Products

Product Documents for Human IL-2 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human IL-2 Antibody

For research use only

Citations for Human IL-2 Antibody

Customer Reviews for Human IL-2 Antibody (2)

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  • Human IL-2 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum
    Species: Cynomolgus Monkey
    Verified Customer | Posted 12/12/2020
    Human IL-2 Antibody AF-202-NA
  • Human IL-2 Antibody
    Name: Zoia Levashova
    Application: Meso Scale Discovery Assay
    Sample Tested: EDTA Plasma
    Species: Mouse
    Verified Customer | Posted 06/19/2018
    Used for detection of IL2 conjugated to the target-specific antibody. Used either as a capture or a detection antibody after labeling with biotin or sulfo-tag, respectively, according to the manufacturer’s protocol (Meso Scale Diagnostics LLC). Detection range of IL-2 conjugates was 10-225,000 pg/ml for anti-IL2-Biotin/anti-Fc gamma-Sulfo-Tag (A) and 50-225,000 pg/ml for anti-Fc gamma-Biotin/anti-IL2-Sulfo-Tag capture/detection combination (B).
    Human IL-2 Antibody AF-202-NA

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Protocols

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